Genotypic effects of calpain 1 and calpastatin on the tenderness of cooked M. longissimus dorsi steaks from Jersey x Limousin, Angus and Hereford-cross cattle.

Single nucleotide polymorphisms (SNPs) in the calpain 1 (CAPN1) and calpastatin (CAST) genes were studied to determine their effects on meat tenderness in Bos taurus cattle. Strip loins (M. longissimus dorsi) were removed from cattle in four resource populations after slaughter (n = 1042), aged under controlled conditions until fixed times after rigor mortis, cooked and measured using a tenderometer. Animals were genotyped for the CAPN1 SNP c.947C>G (p.Ala316Gly; AF252504) and for the CAST SNP c.2959A>G (AF159246). Frequencies of CAPN1 C alleles ranged from 23% to 68%, and CAST A alleles from 84% to 99.5%. From all data combined, the CAPN1 CC genotype (compared with the GG genotype) was associated with a 20.1 +/- 1.7% reduced average shear force at intermediate stages of ageing (P < 0.001) and with a 9.5 +/- 1.3% reduction near ultimate tenderness (P < 0.001). The heterozygote was intermediate. For CAST, corresponding values for AA compared with AG genotypes were reductions of 8.6 +/- 2.0% and 5.1 +/- 1.6% respectively (both P < 0.001), but there were too few GG genotypes for comparison. There were small interactions between the CAPN1 and CAST genotypes. For the CAPN1 and CAST genotypes combined, the maximal genotype effect in average shear force was 25.7 +/- 5.5% (P < 0.001) at intermediate stages and 15.2 +/- 4.8% near ultimate tenderness (P < 0.01).

Understanding the relationship between the inbreeding coefficient and multilocus heterozygosity: theoretical expectations and empirical data.

Geneticists have been interested in inbreeding and inbreeding depression since the time of Darwin. Two alternative approaches that can be used to measure how inbred an individual is involve the use of pedigree records to estimate inbreeding coefficients or molecular markers to measure multilocus heterozygosity. However, the relationship between inbreeding coefficient and heterozygosity has only rarely been investigated. In this paper, a framework to predict the relationship between the two variables is presented. In addition, microsatellite genotypes at 138 loci spanning all 26 autosomes of the sheep genome were used to investigate the relationship between inbreeding coefficient and multilocus heterozygosity. Multilocus heterozygosity was only weakly correlated with inbreeding coefficient, and heterozygosity was not positively correlated between markers more often than expected by chance. Inbreeding coefficient, but not multilocus heterozygosity, detected evidence of inbreeding depression for morphological traits. The relevance of these findings to the causes of heterozygosity--fitness correlations is discussed and predictions for other wild and captive populations are presented.

Bone density in sheep: genetic variation and quantitative trait loci localisation.

Bone density (BD) is an important factor in osteoporotic fracture risk in humans. However, BD is a complex trait confounded by environmental influences and polygenic inheritance. Sheep provide a potentially useful model for studying differences in BD, as they provide a means of circumventing complex environmental factors and are a similar weight to humans. The aims of this study were to establish whether there is genetic variation in BD in sheep and then to localise quantitative trait loci (QTLs) associated with this variation. We also aimed to evaluate the relationship between fat and muscle body components and BD in sheep. Results showed that there was significant (P < 0.01) genetic variation among Coopworth sheep sires for BD. This genetic difference was correlated (P < 0.01) with body weight and muscle mass. A number of QTLs exceeding the suggestive threshold were identified (nine in total). Of these, two (chromosomes 1, P < 0.05; chromosome 24, P < 0.01) were significant using genome-wide permutation significance thresholds (2000 iterations). The position of the QTL on chromosome 24 coincided with a number of other body composition QTLs, indicating possible pleiotropic effects or the presence of multiple genes affecting body composition at that site. This study shows that sheep are potentially a useful model for studying the genetics of BD.

Plasma glucose and insulin levels in genetically lean and fat sheep.

This study investigated whether genetically lean and fat sheep displayed differences in insulin and glucose statuses. Lean genotype sheep had significantly (P < 0.05) greater basal glucose concentrations than fat genotype sheep (4.78 versus 4.52, SED = 0. 104 mmol/l), although basal plasma insulin was not significantly different (mean 304, SEM = 37.3 pmol/l) between the genotypes. During glucose tolerance tests (GTT), carried out at 4 levels of injection: 0, 0.28, 1.39 or 2.78 mmol glucose/kg liveweight, the area under the plasma insulin curve was significantly (P < 0.05) greater for fat than lean genotype sheep, although there were no differences in any glucose parameters. There were no significant differences between genotypes in insulin or glucose concentrations during or following glucose infusion (GINF) experiments at 0, 0.09, 0.46 or 0.93 mmol glucose/kg live-weight/h over 3 hours. Elevated plasma insulin concentrations after a glucose tolerance test are concluded to be associated with increased fatness in this genetically selected line of sheep. However, the differences in insulin and glucose levels between the lean and fat genotype sheep are minor, relative to the differences in carcass composition.

Effects of season, protein and nutritional state on glucose tolerance during an annual cycle of growth in young red deer stags.

Two hypotheses were tested in gonad-intact, young (aged 6-18 months), growing red deer stags during an annual growth cycle. First, that glucose clearance rate is faster during summer than during winter. Secondly, that increased dietary protein availability will enhance winter growth. Stags were randomly assigned into one of two groups: group 1 (n = 5) had 16% while group 2 (n = 6) had 48% of dietary protein naturally protected against fermentative degradation in the rumen. Total crude protein and energy remained similar for each diet (12 and 14% respectively for protein and 11 MJ metabolisable energy/kg dry matter). Stags were kept indoors in individual pens for 12 months and given monthly intravenous glucose tolerance tests (IVGTT), at a dose of 200 mg/kg, in the fed and fasted (48 h) states to determine both growth and steady-state tissue requirements. Protein level had no effect on food intake, weight gain, insulin kinetics, or glucose clearance rate. In the fed state, insulin peak (highest level' after IVGTT) increased (P < 0.01) from October (139 pmol/l) to December (247 pmol/l) (S.E.D. = 42) and remained elevated during the summer, before declining (P < 0.01) from February (223 pmol/l) to April (130 pmol/l) (S.E.D. = 25). Glucose clearance rate was faster (P < 0.05) in December (1.69 litres/min) than June (0.61 litres/min) in the fed state (S.E.D. = 0.30), and decreased (P < 0.05) from February (1.75 litres/min) to April (0.92 litres/min) (S.E.D. = 0.39). During fasting, the pattern of glucose clearance was similar to that observed in the fed state, but the amplitude was lower, while the pattern for insulin peak was similar to that of the fed state. We concluded first, that additional protected protein does not benefit growth during winter. Secondly, we concluded from the fasted, steady-state data that stags are insulin resistant during summer. Thirdly, despite insulin resistance, data on the fed state demonstrated that stags have higher tissue energy requirements during summer growth.

Prolactin does not enhance glucose-stimulated insulin release in red deer stags.

Red deer stags have a seasonal pattern of insulin secretion that is characterized by both elevated basal and glucose-stimulated insulin release in summer compared with winter. Since the seasonal timing of this pattern is similar to that of prolactin and growth rate, the objectives of this study were: first, to determine whether prolactin is associated with the enhanced secretion of insulin during the summer growth period, and second, to determine whether a chronic reduction in plasma prolactin levels would alter body composition. Prolactin was suppressed in plasma using a long-acting form of the dopamine agonist bromocriptine (parlodel LA), which was administered at one of four doses (0-0.3 mg/kg) to each of four groups of castrate stags. Bromocriptine was administered during two 6-wk periods; the first in winter and the second in summer. During the sixth wk of each period, each animal was given three IVGTT at the following glucose doses (10 mg/kg, 70 mg/kg, and 200 mg/kg). Two d later, ovine prolactin was administered to each animal (0.08 mg/kg) and a single IVGTT (70 mg/kg) was given 2 hr later. Body composition was determined by the tritriated water dilution method at the beginning and end of each 6-wk treatment. Chronic suppression of prolactin during winter or summer did not significantly alter the amount of insulin released after each IVGTT, nor did it significantly alter body composition. Furthermore, acute administration of prolactin did not significantly enhance the release of insulin following an IVGTT, during winter or summer treatment periods. It is concluded that elevated levels of prolactin in summer do not enhance the release of insulin to glucose in red deer. Furthermore, a reduction in growth rate following a reduction in plasma prolactin is not associated with a change in body composition.

The physiological effects of natural variation in growth hormone gene copy number in ram lambs.

The effects of natural variation in the number of copies of the growth hormone (GH) gene on growth parameters, plasma GH profiles, and the response to GHRH challenge were compared in Coopworth ram lambs from selection lines differing in body composition and GH levels. Different genotypes at the GH locus carried two, three, or four copies of the GH gene and GH secretion was studied under ad libitum feeding conditions and in the fasted state. There were no significant effects of GH genotype on any parameters of growth or body composition. Basal serum GH concentration, GH pulse frequency, and GH pulse amplitude differed significantly with selection line and fasting, but did not differ significantly between the GH genotypes. Significant differences of subtle nature were found between the GH genotypes in their responsiveness to GHRH. For the ad libitum-fed Lean selection line animals, the first GHRH challenge resulted in a higher mean maximum response for GH1/GH1 than GH2/GH2 (P < 0.05). Between the first and the second challenges there was a decrease in maximum response for the GH1/GH1 genotype and an increase for the GH2/GH2 genotype (P < 0.05 for GH genotype main effect). The differences between GH genotypes in response to GHRH challenge suggest that polymorphism in the number of GH gene copies in sheep may have physiological implications for the function of the GH axis, which may be manifested in growing lambs only under specific genotype-environment combinations.

Seasonal pattern of luteinizing hormone and testosterone pulsatile secretion in young adult red deer stags (Cervus elaphus) and its association with the antler cycle.

Blood from stages aged 15 months (n = 6) was sampled at monthly intervals every 30 min for 24 h for 12 months, at 45 degrees S in New Zealand. Three extra samplings each for 24 h were carried out at about the anticipated time of antler casting. All samples were analysed for luteinizing hormone (LH) and testosterone and the resulting data further analysed by the Pulsar pulse detection routine. The animals were kept indoors under natural daylength and were fed ad libitum. All animals were weighed, antler status and size recorded and testes diameter was measured on each sampling day. Mean LH and testosterone pulsatily and plasma concentration varied seasonally. LH pulse frequency was low during autumn (2.5 pulses in 24 h), winter (1.0-1.5 pulses in 24 h) and early spring (1 pulse in 24 h) and lowest in late spring (0.2 pulse in 24 h) before rising in summer (1.0-4.0 pulses in 24 h). LH pulse amplitude and mean plasma concentration were low (< 1 ng ml-1) from March to November (autumn-spring); both rose to a peak in January (summer) of 3.4 and 1.6 ng ml-1, respectively. Testosterone pulse frequency was generally similar to LH except that slightly more pulses of testosterone than of LH were detected from March to November and more pulses of LH from November to February (summer). Testosterone pulse amplitude fell from March to November (5.3 ng ml-1 to undetectable) although there was a conspicuous peak in July (midwinter) of almost 5 ng ml-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of melatonin implants on secretion of luteinizing hormone in intact and castrated rams.

Rams were treated with melatonin implants in 2 experiments designed to examine the control of reproductive seasonality. In Exp. 1, rams (n = 12) were allocated to 3 treatment groups: 2 groups were treated with 2 melatonin implants per ram for 4 months from 11 November (N) and 9 December (D) and the remaining group was untreated (C). The seasonal increase in luteinizing hormone (LH) pulse frequency and testes size was advanced in Groups N and D. A second seasonal cycle in LH secretion and testes size occurred in Groups N and D after melatonin implants became exhausted. In Exp. 2, rams (n = 20) were allocated to 4 treatment groups: 10 rams were castrated on 6 October and 1 group of entire rams (EM) and one group of castrated rams (CM) were treated with 2 melatonin implants per ram each month from 3 November until 8 January. The other group of entire rams (EC) and castrated rams (CC) was untreated. An increase in LH pulse frequency occurred after castration. Melatonin treatment increased LH pulse frequency in entire rams and reduced LH pulse frequency in castrated rams. The results demonstrated that the advanced reproductive development as a result of treatment with melatonin implants was due to an effect of melatonin on the hypothalamic pulse generator to increase LH pulse frequency. The ability of melatonin to influence LH pulse frequency in entire and castrated rams indicated that an effect of melatonin on the hypothalamic pulse generator is independent of testicular steroids.

Influence of food intake but independence of body weight on puberty in female sheep.

The effect of maintaining female sheep at a body weight intermediate between the normal weight for puberty (30-35 kg) and 20 kg (puberty suppressed) on the onset of oestrous cycles was studied. In addition, the influence of ad-libitum food intake or insulin infusion was studied in animals previously maintained at 20 kg. Coopworth ewe lambs (10 weeks old) were allocated to one of 6 treatments: (A) ad-libitum fed (n = 6), (B) ad-libitum fed to 28 kg then maintained at that weight (n = 6), (C) ad-libitum fed to 24 kg then maintained at that weight (n = 6), (D) maintained at 20 kg until Week 29 and then fed ad libitum (n = 6), (E) maintained at 20 kg and infused with 0.1 U insulin/kg/24 h for 2 weeks from 29-31 weeks of age (n = 5), (F) maintained at 20 kg (n = 6). The lambs were penned indoors under natural photo-period, which was decreasing virtually throughout the study, and fed a pelleted concentrate diet which was recorded daily. They were blood sampled twice a week, and plasma was analysed for progesterone. Puberty was defined as the date when plasma concentrations of progesterone first exceeded 1 ng/ml. In addition, ewes in Groups D, E and F were blood sampled every 10 min for 8 h on Days 0 and +12 of the insulin infusion or access to ad-libitum feeding and the plasma was analysed for luteinizing hormone (LH).(ABSTRACT TRUNCATED AT 250 WORDS)